Enhancement of protein productivity of cell by stabilizing mRNA

通过稳定 mRNA 提高细胞的蛋白质生产率

• 批准号: 03805088
• 负责人: SUZUKI Eiji
• 金额: $ 1.02万
• 依托单位: UNIVERSITY OF TOKYO
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (C)
• 财政年份: 1991
• 资助国家: 日本
• 起止时间: 1991 至 1992
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-03805088/
• 关键词: Stability of mRNA; Rate of protein synthesis; Animal cell; Cell culture; Production of protein; Modification of mRNA; Messenger ribonucleic acid; Rate of decomposition; 蛋白質生合成速度; mRNAの改造; メッセンジャーリボ核酸の安定性; メッセンジャ-リボ核酸の安定性

A structured model predicts that stabilization of mRNA combined with suppression of cell growth enhances the protein production rate of mammalian cells. We confirmed this prediction experimentally as follows.1. Choosing antibody as an example of ehich the mRNA was stable we confirmed that the mRNA accumulated in cells and the protein production rate per cell increased at the same time as the growth rate of the cells decreased.3. Addition of caffeine or interleukin 6 into culture medium was the best method to suppress cell growth rate for enhancing the protein productivity.2. We tried to stabilize mRNA of mouse interleukin 2 which is an example of unstable mRNAs by deleting AU rich region at the 3' untranslated region. Mouse myeloma cells transfected with the modified mRNA produced enhanced amount of the interleukin.

一个结构化的模型预测，mRNA的稳定和细胞生长的抑制相结合，提高了哺乳动物细胞的蛋白质产量。我们在实验上证实了这一预测，如下所示。以抗体为例，当细胞中的mRNA稳定时，证实了随着细胞生长速度的降低，细胞内的mRNA积累增加，单位细胞的蛋白质产量也随之增加。结论：1.在培养基中添加咖啡因或白介素6是抑制细胞生长速度、提高蛋白质产量的最好方法。我们试图通过删除3‘非翻译区的AU富集区来稳定小鼠白细胞介素2的mRNA，这是一个不稳定的mRNAs的例子。转基因小鼠骨髓瘤细胞产生较高水平的白介素量。

项目成果

Satoshi Terada: "Molecular Biological Analysis of Perfusion Culture of Hybridoma and Effective Production of Monoclonal Antibody Using Interleukin-6" KAGAKU KOGAKU RONBUNSHU. 19. 207-213 (1993)

Satoshi Terada：“杂交瘤灌注培养的分子生物学分析和使用白细胞介素6有效生产单克隆抗体”KAGAKU KOGAKU RONBUNSHU。

Fusao Makishima: "Interleukinー6 is antiproliferative to a mouse hybridoma cell line and promotive for its antibody productivity" Cytotechnology. (1992)

Fusao Makishima：“Interleukin-6 对小鼠杂交瘤细胞系具有抗增殖作用，并促进其抗体生产力”Cytotechnology (1992)。

鈴木栄二: "細胞増殖抑制によるハイブリドーマの抗体生産性向上 : 最適増殖抑制方法のモデルと実験による探索" 化学工学論文集. 19. 198-206 (1993)

Eiji Suzuki：“通过抑制细胞增殖提高杂交瘤的抗体生产力：使用模型和实验寻找最佳的增殖抑制方法”《化学工程杂志》19. 198-206 (1993)。

E.Suzuki: "A SIMPEE STRUCTURED MODEL PREDICTED POSITIVELYー,NEGATIVELYOR NONーGROWTH ASSOCIATED ANTIBODY PRODUCTION RATE DEPENDING ON CULTURE CONDITIONS" Proceedings of the Fourth Annual Meeting of Japanese Association for Animal Cell Technology. (1992)

E.Suzuki：“根据培养条件预测正向、负向或非生长相关抗体产生率的 SIMPEE 结构模型”日本动物细胞技术协会第四届年会记录（1992 年）。

鈴木 栄二: "細胞増殖抑制によるハイブリドーマの抗体生産性向上:最適増殖抑制方法のモデルと実験による探索" 化学工学論文集. (1993)

Eiji Suzuki：“通过抑制细胞增殖提高杂交瘤的抗体生产力：使用模型和实验寻找最佳的增殖抑制方法”《化学工程杂志》（1993）。